Journal: Open Life Sciences

Article Title: Sini Decoction Intervention on Atherosclerosis via PPARγ-LXRα-ABCA1 Pathway in Rabbits

doi: 10.1515/biol-2018-0053

Figure Lengend Snippet: Expression profiling of PPARγ-LXRα signaling to confirm the mechanism of treating AS by SND. The PPARγ (A-D) and LXRα (E-H) proteins levels in liver, peritoneal macrophages, PMC, and adipose tissue were measured by western blotting. The relative quantities were analyzed by ImageJ software. Data shown as mean ± SEM (n = 6). # P<0.05, compared with Chow group; *P<0.05, compared with HFD group.

Article Snippet: Primary antibodies to ABCA1 and Apo-B were purchased from Abcam (MA, USA); PPARγ antibody and LXRα antibody were purchased from Proteintech (IL, USA); and ApoA-I antibody was purchased from MyBioSource (CA, USA).

Techniques: Expressing, Western Blot, Software

Journal: Drug Design, Development and Therapy

Article Title: Asiatic acid protests against myocardial ischemia/reperfusion injury via modulation of glycometabolism in rat cardiomyocyte

doi: 10.2147/DDDT.S175116

Figure Lengend Snippet: Asiatic acid increases PPARγ levels and promotes GLUT4 translocation to plasma membrane in ischemic rats. Notes: The myocardium was isolated from vehicle-, asiatic acid (AA)-, and asiatic acid plus LY294002 (AA+LY)-treated rats under normal condition and following 1 hour of myocardial ischemia and 24 hours of reperfusion, respectively. The mRNA levels of ( A ) PPARγ and ( B ) GLUT4 were determined by real-time PCR. ( C ) PPARγ, GLUT4 from plasma membrane (PM GLUT4), GLUT4, and GAPDH levels were determined by Western blot. ( D ) Quantitative analysis of PPARγ levels were normalized to GAPDH levels, and ( E ) GLUT4 translocation levels (PM GLUT4) were normalized to total GLUT4. # P <0.05, ## P <0.01 vs the sham group; ** P <0.01 vs the vehicle group; && P <0.01 vs the asiatic acid-treated group. Abbreviation: MI/R, myocardial ischemia/reperfusion.

Article Snippet: The anti-Akt and anti-phosphor-Akt antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA); the anti-GSK-3β and anti-phospho-GSK-3β (Ser9) antibodies were from Cell Signaling Technology, Inc. (Danvers, MA, USA); the anti-GLUT4 and anti-PPARγ antibody was from Boster Biological Technology, Ltd. (Wuhan, China); and the anti-glyceraldehyde phosphate dehydrogenase (GAPDH) antibody was obtained from KangChen Bio-tech Inc. (Shanghai, China).

Techniques: Translocation Assay, Clinical Proteomics, Membrane, Isolation, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 1. Treatment with PPAR agonists or overexpression of PPARs downregulated the mRNA and protein expression levels of CRBN. (A). Pathway enrichment analysis based on the microarray assay in MM cells treated with PIM kinase inhibitors; red arrows indicated PPAR pathway. (B). MM1.R, NCIH929 and RPMI8226 cells lines were treated with 5 μM or the EC50 concentration of individual PPAR agonist for 48 h, and levels of CRBN mRNA were measured by qRT-PCR. (C). MM1.R, NCIH929 and RPMI8226 cells were treated with 5 μM or the EC50 concentration of individual PPAR agonist for 48 h, and CRBN and GAPDH protein expression levels were assessed by Western blot. Bar graph represents the ratio of CRBN/GAPDH protein. One of 3 independent experiments was shown. (D). Representative images of DAPI and CRBN staining in NCIH929 cells treated with 5 μM of PPAR agonists for 48 h. (E). NCIH929 and MM1.R cells were transfected with a Lenti-PPARα, Lenti- PPARβ/δ, or Lenti-PPARγ expressing plasmid or control vector for 48 h. CRBN protein expression level was measured by Western blot.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Over Expression, Expressing, Microarray, Concentration Assay, Quantitative RT-PCR, Western Blot, Staining, Transfection, Plasmid Preparation, Control

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 2. PPAR agonists reduce the anti-myeloma activity of lenalidomide in vitro. (A).

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Activity Assay, In Vitro

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 3. Overexpression of PPAR resulted in reduced activity of lenalidomide in vitro. (A). NCIH929 and RPMI8226 cell lines were transfected with control vector or a PPAR over-expressing plasmid for 24 h and then treated with lenalidomide for additional 48 h. Cell viability was measured by MTT assay. (B). NCIH929 and RPMI8226 cell lines were transfected with control vector or a CRBN over-expressing plasmid for 24 h and then treat with lenalidomide alone or in combination with PPAR agonists for 48 h. Cell viability was measured by MTT assay. (C). NCIH929 and RPMI8226 cells were transduced with CRBN specific CRISP/cas9 knockout plasmid for 24 h and then treated with lenalidomide, PPAR agonists alone, or in com bination for additional 48 h. Cell viability was measured by MTT assay. Results are presented as mean ± SD from at least three separate experiments. *: p < 0.05; **: p < 0.01; ***: p < 0.001.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Over Expression, Activity Assay, In Vitro, Transfection, Control, Plasmid Preparation, Expressing, MTT Assay, Transduction, Knock-Out

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 4. PPAR agonists reduce the anti-myeloma ac tivity of lenalidomide in vivo. (A). Bioluminescence intensity showing the different tumor burdens in NSG mice implanted with MM1.R myeloma cells and treated with control PBS (CTL), fenofibrate (Fen), lenalidomide (Len), or the combination (Len and Fen) (7 mice per group) from week 1 to week 10. (B). Left panel: the bioluminescence activity was quantified by determining the total flux (photons/sec) in each mouse from week 1 to week 10. Right panel: repre sentative images of tumors harvested from the control group (CTL) and the fenofibrate group (Fen) at week 8 and tumors harvested from the lenalidomide group (Len) and the combination group (Len and Fen) at week 10. (C). CRBN mRNA expression of the har vested tumors. qRT-PCR was used to detect the CRBN mRNA expression in the harvested tumors.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: In Vivo, Control, Activity Assay, Expressing, Quantitative RT-PCR

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 5. PPARs directly regulate transcription of CRBN in MM cells. (A). Representative images of ChiP-PCR products amplified by primers (100bp) on 2% agarose gel in NCIH929 cells. (B). Bar graph of ChiP DNA shows fold change of CRBN promoter binding after treatment with PPAR agonists. (C). NCIH929 cells were transfected with CRBN/PGL3 firefly luciferase reporter vector construct. The cells were then treated with PPAR agonists and luciferase bio-luminate activity was measured. (D). NCIH929 cells were transfected with mutated CRBN/PGL3 firefly luciferase reporter vector construct. The cells were then treated with PPAR agonists and luciferase bio-luminate activity was measured.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Amplification, Agarose Gel Electrophoresis, Binding Assay, Transfection, Luciferase, Plasmid Preparation, Construct, Activity Assay

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 6. Co-administration of PPAR antagonists in crease CRBN expression and improve the sensitivity to lenalidomide. (A) NCIH929 cells were transfected with CRBN/PGL3 firefly luciferase reporter vector construct. The cells were then treated with PPAR antagonists and luciferase bio-luminate activity was measured. (B) MM1.R and NCIH929 cells were treated with PPAR antagonists (GW6471 10 μM, GSK3787 10 μM and GW9662 10 μM) or DMSO control for 48 h. CRBN expression was then measured by Western blot analysis. (C). MM1.R and NCIH929 were treated DMSO, lenalidomide, PPAR antagonists, or in combination for 48 h. Cell viability was measured by MTT assays. Results are presented as mean ± SD from at least three separate experiments. *: p < 0.05; **: p < 0.01; ***: p < 0.001.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Expressing, Transfection, Luciferase, Plasmid Preparation, Construct, Activity Assay, Control, Western Blot

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 7. PPAR expression is upregulated in MM patients and is associated with poorer clinical outcomes. (A). Bone marrow biopsy sections from newly diagnosed MM patients and normal controls were stained with PPAR antibodies. Representative images of PPAR staining were shown. (B). Box plot shows the IHC scores of PPARs expression in CD138+ and CD138-cells in bone marrow biopsy samples of newly diagnosed myeloma patients and in normal control bone marrows. (C) Progression free survival (left) and overall survival (right) between newly diagnosed myeloma patients with high total PPAR IHC score (IHC score >8) and newly diagnosed myeloma patients with low total PPAR IHC score (IHC score ≤4). (D). Progression free survival and overall survival between patients with high PPARβ/δ expression (IHC >4) and patients with low PPARβ/δ expression (IHC ≤4). (E). Progression free survival and overall survival between patients with high PPARγ expression (IHC >4) and patients with low PPARγ expression (IHC ≤4). IHC scores represented the sum of score for CD138+ cells and score for CD138-cells.

Article Snippet: PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC (Littleton, CO).

Techniques: Expressing, Staining, Control